Compositions of statins and omega-3 fatty acids

ABSTRACT

The present disclosure provides pharmaceutical compositions of statins and polyunsaturated fatty acids (PUFAs), in which the statins are dissolved in the PUFAs, the PUFA species being present substantially in the free acid form. Also provided are oral unit dosage forms of the disclosed pharmaceutical compositions and methods of treating blood lipid disorders using the compositions and oral unit dosage forms.

1. CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit under 35 U.S.C. §119(e) of U.S.Provisional Application No. 61/643,764, filed May 7, 2012, the contentof which is incorporated herein by reference in its entirety.

2. BACKGROUND

Pharmaceutical compositions rich in omega-3 polyunsaturated fatty acids(PUFAs) have been developed to treat a variety of clinical indications,including various disorders of blood lipids, includinghypertriglyceridemia and mixed dyslipidemias. Statin monotherapy iswidely used to treat hypercholesterolemia and other blood lipiddisorders.

Numerous studies have reported on the increased benefits of combinedtreatment with statins and omega-3 PUFAs in various cardiovasculardisorders (See Nakamura et al., 1999, Int. J. Clin. Lab Res. 29:22-25and Davidson et al., 1997, Am J Cardiol 80:797-798). To improve patientconvenience and compliance, dual compositions comprising statins andomega-3 PUFAs in esterified form have been described, but some statinsare known to be poorly soluble in such formulations (see, e.g., U.S.Pat. No. 7,642,287), and no such dual composition is commerciallyavailable.

Therefore, there is a need in the art for compositions that combinestatins and omega-3 PUFAs in a single oral unit dosage form, and thatprovide enhanced solubility and bioavailability. Such pharmaceuticalcompositions would improve patient convenience, and may allow fortreatment with improved effectiveness, fewer excipients, and betterpatient compliance, than is provided by the administration of the twoactives in separate unit dosage forms.

3. SUMMARY

The inventors have discovered that certain statins have unexpectedlygreater solubility in a PUFA free acid solvent system, a PUFAcomposition in which each of the PUFA species is present substantiallyin the free acid form, as compared to their solubility in a PUFA ethylester solvent system, a PUFA composition in which each of the PUFAspecies is present substantially in esterified form. Accordingly, thepresent disclosure provides pharmaceutical compositions, oral unitdosage forms, dosage kits, and methods of treatment comprising at leastone statin dissolved in a PUFA free acid solvent system. In typicalembodiments, the statin is rosuvastatin. In some embodiments, the statinis atorvastatin, pravastatin, lovastatin, simvastatin, atorvastatin,fluvastatin, or pitavastatin.

In a first aspect, pharmaceutical compositions comprising at least onestatin dissolved in a PUFA free acid solvent system are provided.

In various embodiments, at least 90% of the total polyunsaturated fattyacid in the PUFA free acid solvent system is present in the free acidform. In certain embodiments, at least 91%, at least 92%, at least 93%,at least 94%, at least 95%, at least 96%, at least 97%, at least 98%,even at least 99% of the total polyunsaturated fatty acid in the PUFAfree acid solvent system is present in the free acid form.

In typical embodiments, the PUFA free acid solvent system comprises atleast one, typically a plurality of, omega-3 PUFA species, wherein eachof the omega-3 PUFA species is present substantially in the free acidform. In various embodiments, the composition comprises EPA, DHA, andDPA (n-3), each substantially in free acid form.

In some embodiments, at least 80% by weight of the statin is dissolvedin the PUFA free acid solvent system (less than 20% by weight of thestatin is undissolved in the solvent system), while in particularembodiments, at least 90% by weight of the statin is dissolved in thePUFA free acid solvent system (less than 10% by weight of the statin isundissolved in the solvent system).

In various embodiments, the one or more statins is dissolved in the PUFAfree acid solvent system in an amount that permits administration of atherapeutic amount of statin in a convenient number of oral unit doses,which number of oral unit doses also suffices to deliver a therapeuticdose of omega-3 PUFAs.

In certain embodiments, the one or more statins is present in thepharmaceutical composition in an amount, per ml or per g of PUFA freeacid solvent system, of at least about 5 mg, at least about 10 mg, atleast about 15 mg, at least about 20 mg, at least about 25 mg, at leastabout 30 mg, at least about 40 mg, at least about 50 mg, at least about60 mg, at least about 70 mg, at least about 80 mg, at least about 90 mg,even at least about 100 mg, at least about 110 mg, and in certainembodiment, at least about 120 mg.

In selected embodiments, one or more statins is present in thepharmaceutical composition in an amount, per ml or per g of PUFA freeacid solvent system, of from about 2 mg to about 80 mg, from about 5 mgto about 60 mg, or from about 10 mg to about 40 mg, or from about 20 mgto about 30 mg. In various embodiments, the at least one statin isselected from the group consisting of rosuvastatin, pravastatin,lovastatin, simvastatin, atorvastatin, fluvastatin, and pitavastatin.

In another aspect, oral unit dosage forms comprising the pharmaceuticalcompositions of the disclosure are provided. In some embodiments, thepharmaceutical composition is encapsulated in a soft gelatin capsule. Invarious embodiments, the oral unit dosage form further comprises atleast one coating on the exterior of the soft gelatin capsule. In someembodiments, each of the at least one coatings is selected from thegroup consisting of: cellulose acetate trimellitate, cellulose acetatephthalate and poly(ethylacrylate-methylacrylate).

In some embodiments, the soft gelatin capsule encapsulates 1-100 mg of astatin dissolved in about 250 mg, about 500 mg, or about 1000 mg of aPUFA free acid solvent system. In certain embodiments, the soft gelatincapsule encapsulates 2-40 mg of a statin dissolved in about 250 mg,about 500 mg, or about 1000 mg of a PUFA free acid solvent system.

In other aspects, the disclosure provides methods of treating a bloodlipid disorder, comprising administering an effective amount of apharmaceutical composition of the disclosure. The blood lipid disorderis selected, in certain embodiments, from the group consisting of:hypertriglyceridemia, hypercholesterolemia, coronary heart disease(CHD), hypertriglyceridemia, mixed dyslipidemias, heart failure,myocardial infarction, cardiac arrhythmias, ischemic dementia,hypertension, nephropathy, retinopathy, vascular disease, andatherosclerotic disease.

4. BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 provides data showing solubility of atorvastatin calcium invarious pH buffers; and

FIG. 2 provides data showing solubility of rosuvastatin calcium invarious pH buffers.

5. DETAILED DESCRIPTION 5.1. Overview

Statins are known to be poorly soluble in compositions of omega-3 PUFAsin which the PUFAs are present substantially in esterified form (“PUFAethyl ester solvent systems”). See, e.g., U.S. Pat. No. 7,642,287. Thepresent inventors have now discovered that certain statins unexpectedlyhave significantly greater solubility in PUFA compositions in which eachof the PUFA species is present substantially in the free acid form(“PUFA free acid solvent system”), as compared to their solubility in aPUFA ethyl ester solvent system. Accordingly, the present disclosureprovides pharmaceutical compositions, oral unit dosage forms, and dosagekits comprising at least one statin dissolved in a PUFA free acidsolvent system, and methods of using these compositions and unit oraldosage forms for treatment of blood lipid disorders. In typicalembodiments, the statin is rosuvastatin. In some embodiments, the statinis atorvastatin, pravastatin, lovastatin, simvastatin, atorvastatin,fluvastatin, or pitavastatin.

5.2. Pharmaceutical Compositions

In a first aspect, pharmaceutical compositions comprising at least onestatin dissolved in a PUFA free acid solvent system are provided. Intypical embodiments, the PUFA free acid solvent system comprises atleast one, typically a plurality of, omega-3 PUFA species, wherein eachof the omega-3 PUFA species is present substantially in the free acidform. In some embodiments, the statin is rosuvastatin. In variousembodiments, the statin is atorvastatin, pravastatin, lovastatin,simvastatin, atorvastatin, fluvastatin, or pitavastatin.

In some embodiments of the pharmaceutical composition, the statin isdissolved in the PUFA free acid solvent system to provide asubstantially homogeneous composition.

In some embodiments, the statin is completely dissolved in the PUFA freeacid solvent system. In various embodiments, the statin is substantiallydissolved (i.e., at least about 90% of the statin is dissolved in thePUFA free acid solvent system, with about 10% or less of the statin isundissolved in the PUFA free acid solvent system). In certainembodiments, at least about 70% of the statin is dissolved in the PUFAfree acid solvent system. In some embodiments, at least about 80%, 85%,90%, 95%, even at least about 96%, 97%, 98%, 99%, even at least about99.5% of the statin is dissolved in the PUFA free acid solvent system.In some embodiments, the statin is less than 30%, less than 20%, lessthan 15%, less than 10%, or less than 5% undissolved in the PUFA freeacid solvent system.

In some embodiments, the pharmaceutical composition does not requiresignificant amounts of compounds other than polyunsaturated fattyacids—such as surfactants, hydrophilic or hydrophobic solvents, oils orcombinations thereof (individually and collectively, “non-PUFAsolubilizers”)—to dissolve the statin in the PUFA free acid solventsystem.

Preferably, the statin is dissolved in the pharmaceutical compositionwithout the use of any non-PUFA solubilizers.

If present, non-PUFA solubilizers are present in amounts of 50% or less(w/w) based on the total weight of the PUFA free acid solvent system,40% or less, 30% or less, 20% or less, 10% or less, or 5% or less. Invarious embodiments, the weight ratio of polyunsaturated fatty acids tonon-PUFA solubilizers is at least about 1:1, at least 5:1, or at least10:1. In some embodiments, the PUFA free acid solvent system contains nonon-PUFA solubilizers.

Various embodiments of the pharmaceutical composition comprise a statindissolved in a PUFA free acid solvent system, wherein the statin ispresent in the PUFA free acid solvent system in a weight-to-volume (w/v)ratio of about 1:5,000; about 1:4,500; about 1:4,000; about 1:3,50;about 1:3,000; about 1:2,500; about 1:2,000; about 1:1,500; about1:1,000; about 1:800; about 1:700; about 1:500; about 1:400: about1:300; about 1:250, about 1:125, about 1:100; about 1:80; about 1:50; orratios intermediate the above-recited values.

In a variety of embodiments, at least one statin is dissolved in thePUFA free acid solvent system, wherein the statin is present in aweight-to-weight (w/w) ratio with respect to the PUFA free acid solventsystem of about 1:5,000; about 1:4,500; about 1:4,000; about 1:3,500;about 1:3,000; about 1:2,500; about 1:2,000; about 1:1,500; about1:1,000; about 1:800; about 1:700; about 1:500; about 1:400: about1:300; about 1:250, about 1:125, about 1:100; about 1:80; about 1:50, orratios intermediate these values.

In certain embodiments, the pharmaceutical composition comprises astatin dissolved in a PUFA free acid solvent system, wherein the statinis dissolved in the PUFA free acid solvent system at a concentration ofat least about 0.1 mg/ml solvent system, 0.2 mg/ml, 0.5 mg/ml, 1 mg/ml,2 mg/ml, 5 mg/ml, 7.5 mg/ml, 10 mg/ml, 15 mg/ml, 20 mg/ml, 30 mg/ml, 40mg/ml, 50 mg/ml, 75 mg/ml, 100 mg/ml, 120 mg/ml, 150 mg/ml, or 200mg/ml, or in concentrations intermediate these values. In variousembodiments, the pharmaceutical composition comprises a statin dissolvedin a PUFA free acid solvent system, wherein the statin is dissolved inthe PUFA free acid solvent system at a concentration of at least about0.1 mg/g solvent system, 0.2 mg/g solvent system, 0.5 mg/g solventsystem, 1 mg/g solvent system, 2 mg/g solvent system, 5 mg/g solventsystem, 7.5 mg/g solvent system, 10 mg/g solvent system, 15 mg/g solventsystem, 20 mg/g solvent system, 30 mg/g solvent system, 40 mg/g solventsystem, 50 mg/g solvent system, 75 mg/g solvent system, 100 mg/g solventsystem, 120 mg/g solvent system, 150 mg/g solvent system, or 200 mg/gsolvent system, or in concentrations intermediate these values.

5.2.1. PUFA Free Acid Solvent System

The PUFA free acid solvent system comprises at least one, typically aplurality of, species of omega-3 PUFAs, each present substantially inthe free acid form.

In typical embodiments, the PUFA free acid solvent system compriseseicosapentaenoic acid (C20:5 n-3) (“EPA,” also known as timnodonic acid)substantially in free acid form. In various embodiments, the PUFA freeacid solvent system comprises docosahexaenoic acid (C22:6 n-3) (“DHA,”also known as cervonic acid) substantially in free acid form. Inselected embodiments, the PUFA free acid solvent system comprisesdocosapentaenoic acid (C22:5 n-3) (“DPA,” also known as clupanodonicacid), substantially in free acid form.

In characteristic embodiments, the PUFA free acid solvent systemcomprises EPA, substantially in free acid form, in an amount, calculatedas a percentage by area on GC chromatogram of all fatty acids in thePUFA free acid solvent system, of at least about 40% (“40% (a/a)”). Invarious embodiments, the PUFA free acid solvent system comprises EPA inan amount of at least about 41% (a/a), 42% (a/a), 43% (a/a), 44% (a/a),45% (a/a), 46% (a/a), 47% (a/a), 48% (a/a), 49% (a/a), or at least about50% (a/a). In certain embodiments, the PUFA free acid solvent systemcomprises EPA, substantially in free acid form, in an amount of at leastabout 51% (a/a), at least about 52% (a/a), at least about 53% (a/a), atleast about 54% (a/a), at least about 55% (a/a), at least about 56%(a/a), at least about 57% (a/a), at least about 58% (a/a), even at leastabout 59% (a/a), at least about 60% (a/a), at least about 61% (a/a), 62%(a/a), 63% (a/a), 64% (a/a), or 65% (a/a).

In a variety of embodiments, the PUFA free acid solvent system comprisesEPA, substantially in free acid form, in an amount of at least about 70%(a/a), at least about 75% (a/a), at least about 80% (a/a), at leastabout 85% (a/a), 90% (a/a), 91% (a/a), 92% (a/a), 93% (a/a), 94% (a/a),even at least about 95% (a/a), 96% (a/a), 97% (a/a), 98% (a/a), or 99%(a/a).

In certain embodiments, the PUFA free acid solvent system comprises EPA,substantially in free acid form, in an amount of about 45 to about 65%(a/a). In particular embodiments, EPA is present, substantially in freeacid form, in an amount of about 50 to about 60% (a/a). In variousembodiments, EPA is present, substantially in free acid form, in anamount of about 52 to about 58.0% (a/a). In some embodiments, EPA ispresent, substantially in free acid form, in an amount of about 55%(a/a) to about 56% (a/a). In some embodiments, EPA is present in anamount of about 55% (a/a).

In various embodiments, the PUFA free acid solvent system comprises EPA,substantially in free acid form, in an amount, calculated as apercentage by mass of all fatty acids in the PUFA free acid solventsystem (“% (m/m)”), of about 50% (m/m) to about 60% (m/m). In certainembodiments, EPA is present, substantially in free acid form, in anamount of about 55% (m/m).

In certain embodiments, the PUFA free acid solvent system comprises DHA,substantially in free acid form, in an amount of at least about 13%(a/a). In various embodiments, the PUFA free acid solvent systemcomprises DHA, substantially in free acid form, in an amount of at leastabout 14% (a/a), at least about 15% (a/a), at least about 16% (a/a), atleast about 17% (a/a), at least about 18% (a/a), at least about 19%(a/a), or at least about 20% (a/a). In selected embodiments, the PUFAfree acid solvent system comprises DHA, substantially in free acid form,in an amount of at least about 21% (a/a), at least about 22% (a/a), atleast about 23% (a/a), at least about 24% (a/a), even at least about 25%(a/a).

In a variety of embodiments, the PUFA free acid solvent system comprisesDHA, substantially in free acid form, in an amount of at least about 30%(a/a), 35% (a/a), even at least about 40% (a/a).

In various embodiments, the PUFA free acid solvent system comprises DHA,substantially in free acid form, in an amount of about 13% (a/a) toabout 25% (a/a). In certain embodiments, DHA is present, substantiallyin free acid form, in an amount of about 15% (a/a) to about 25% (a/a).In several embodiments, DHA is present, substantially in free acid form,in an amount of about 17% (a/a) to about 23% (a/a). In certainembodiments, DHA is present, substantially in free acid form, in anamount of about 19% (a/a) to about 20% (a/a).

In various embodiments, the PUFA free acid solvent systems comprise DHA,substantially in free acid form, in an amount of about 15% (m/m) toabout 25% (m/m). In certain embodiments, DHA is present, substantiallyin free acid form, in an amount of about 17% (m/m) to about 23% (m/m).In certain embodiments, DHA is present, substantially in free acid form,in an amount of about 20% (m/m).

In certain embodiments, the PUFA free acid solvent system comprises DPA,substantially in free acid form, in an amount of at least about 1%(a/a). In various embodiments, the PUFA free acid solvent systemcomprises DPA, substantially in free acid form, in an amount of at leastabout 1.5% (a/a), 2% (a/a), 2.5% (a/a), 3% (a/a), 3.5% (a/a), 4% (a/a),4.5% (a/a), even at least about 5% (a/a). In selected embodiments, thePUFA free acid solvent system comprises DPA, substantially in free acidform, in an amount of at least about 6% (a/a), at least about 7% (a/a),at least about 8% (a/a), or at least about 9% (a/a).

In a variety of embodiments, the PUFA free acid solvent system comprisesDPA, substantially in free acid form, in an amount of about 1% (a/a) toabout 8% (a/a). In certain embodiments, the PUFA free acid solventsystem comprises DPA, substantially in free acid form, in an amount ofabout 2% (a/a) to about 7% (a/a). In selected embodiments, the PUFA freeacid solvent system comprises DPA, substantially in free acid form, inan amount of about 3% (a/a) to about 6% (a/a). In particularembodiments, the PUFA free acid solvent system comprises DPA,substantially in free acid form, in an amount of about 4% (a/a) to about5% (a/a).

In various embodiments, the PUFA free acid solvent system comprises DPA,substantially in free acid form, in an amount of no less than about 1%(m/m). In various embodiments, the PUFA free acid solvent systemcomprises DPA, substantially in free acid form, in an amount of about 1%(m/m) to about 8% (m/m). In particular embodiments, the PUFA free acidsolvent system comprises DPA, substantially in free acid form, in anamount of no more than about 10% (m/m).

In a variety of embodiments, the PUFA free acid solvent system comprisesEPA and DHA, each substantially in free acid form.

In various embodiments, the PUFA free acid solvent system comprises EPAand DHA, each substantially in free acid form, in a total amount of atleast about 60% (a/a). In various embodiments, the PUFA free acidsolvent system comprises EPA and DHA, each substantially in free acidform, in a total amount of at least about 61% (a/a), 62% (a/a), 63%(a/a), 64% (a/a), 65% (a/a), 66% (a/a), 67% (a/a), 68% (a/a), 69% (a/a),or at least about 70% (a/a). In particular embodiments, the PUFA freeacid solvent system comprise EPA and DHA, each substantially in freeacid form, in a total amount of at least about 71% (a/a), 72% (a/a), 73%(a/a), 74% (a/a), 75% (a/a), 76% (a/a), 77% (a/a), 78% (a/a), 79% (a/a),even at least about 80% (a/a). In certain embodiments, the PUFA freeacid solvent system comprises EPA and DHA, each substantially in freeacid form, in total amount of at least about 81% (a/a), 82% (a/a), atleast about 83% (a/a), 84% (a/a), even at least about 85% (a/a).

In various embodiments, the PUFA free acid solvent system comprises EPAand DHA, each substantially in free acid form, in a total amount ofabout 70% (m/m) to about 80% (m/m). In certain embodiments, the PUFAfree acid solvent system comprises about 75% (m/m) EPA plus DHA, eachsubstantially in free acid form.

In certain embodiments, the PUFA free acid solvent system comprises EPAand DHA, each substantially in free acid form, in mass ratios of atleast about 2:1, 2.1:1, 2.2:1, 2.3:1, 2.4:1, 2.5:1, 2.6:1, 2.7:1, 2.8:1,2.9:1, even at least about 3:1. In certain embodiments, the PUFA freeacid solvent system comprises EPA and DHA, each substantially in freeacid form, in a mass ratio of about 2.75:1.

In various embodiments, the PUFA free acid solvent system comprises EPA,DHA, and DPA, each substantially in free acid form, in a total amount ofat least about 61% (a/a). In typical embodiments, the PUFA free acidsolvent system comprises EPA, DHA, and DPA, each substantially in freeacid form, in a total amount of at least about 62% (a/a), 63% (a/a), 64%(a/a), 65% (a/a), 66% (a/a), at least about 67% (a/a), at least about68% (a/a), at least about 69% (a/a), or at least about 70% (a/a). Incertain embodiments, the PUFA free acid solvent system comprises EPA,DHA, and DPA, each substantially in free acid form, in a total amount ofat least about 71% (a/a), 72% (a/a), 73% (a/a), 74% (a/a), 75% (a/a),76% (a/a), 77% (a/a), 78% (a/a), 79% (a/a), 80% (a/a), even at leastabout 81% (a/a), 82% (a/a), 83% (a/a), 84% (a/a), 85% (a/a), 86% (a/a),87% (a/a), even at least about 88% (a/a).

In various embodiments, the PUFA free acid solvent system comprises EPA,DHA, and DPA, each substantially in free acid form, in a total amount ofbetween about 70% (a/a) to about 90% (a/a).

In a particular series of embodiments, EPA, substantially in free acidform, is present in an amount of about 55% (a/a) to about 56% (a/a);DHA, substantially in free acid form, is present in an amount of about19% (a/a) to about 20% (a/a); and DPA, substantially in free acid form,is present in an amount of about 4% (a/a) to about 5% (ala). In certainembodiments, EPA, substantially in free acid form, is present in anamount of about 55% (m/m) to about 56% (m/m); DHA, substantially in freeacid form, is present in an amount of about 19% (m/m) to about 20%(m/m); and DPA, substantially in free acid form, is present in an amountof about 4% (m/m) to about 5% (m/m).

In certain embodiments, the PUFA free acid solvent system is a complexmixture comprising a plurality of species of omega-3 PUFAs and aplurality of species of omega-6 PUFAs, each present substantially infree acid form.

In certain embodiments, the PUFA free acid solvent system comprises EPA,DHA, DPA, and further comprises one or more omega-3 polyunsaturatedfatty acid species selected from the group consisting of α-linolenicacid (C18:3 n-3), moroctic acid (C18:4 n-3, also known as stearidonicacid), eicosatrienoic acid (C20:3 n-3), eicosatetraenoic acid (C20:4n-3), and heneicosapentaenoic acid (C21:5 n-3), each substantially inthe free acid form.

In particular embodiments, the PUFA free acid solvent system comprisesEPA, DHA, DPA, and moroctic acid, each substantially in the free acidform. In a variety of embodiments, the PUFA free acid solvent systemcomprises EPA, DHA, DPA, moroctic acid, and heneicosapentaenoic acid,each substantially in the free acid form. In specific embodiments, thePUFA free acid solvent system comprises EPA, DHA, DPA, moroctic acid,heneicosapentaenoic acid, and eicosatetraenoic acid, each substantiallyin the free acid form. In selected embodiments, the PUFA free acidsolvent system comprises EPA, DHA, DPA, α-linolenic acid (C18:3 n-3),moroctic acid (C18:4 n-3), eicosatrienoic acid (C20:3 n-3),eicosatetraenoic acid (C20:4 n-3), and heneicosapentaenoic acid (C21:5n-3), each substantially in the free acid form.

In various embodiments, total omega-3 fatty acids—defined as the sum ofalpha-linolenic acid (C18:3 n-3), moroctic acid (C18:4 n-3),eicosatrienoic acid (C20:3 n-3), eicosatetraenoic acid (C20:4 n-3),eicosapentaenoic acid (EPA) (C20:5 n-3), heneicosapentaenoic acid (C21:5n-3), docosapentaenoic acid (C22:5 n-3) and docosahexaenoic acid (DHA)(C22:6 n-3), each present substantially in the free acid form—constitutefrom about 80% (a/a) to about 95% (a/a) of all fatty acids in the PUFAfree acid solvent system. In a variety of embodiments, total omega-3fatty acids constitute from about 80-about 95% (m/m) of all fatty acidsin the PUFA free acid solvent system.

In various embodiments, the PUFA free acid solvent system furthercomprises one or more species of omega-6 PUFA, each presentsubstantially in the free acid form.

In certain embodiments, the PUFA free acid solvent system comprises oneor more species of omega-6 PUFA selected from the group consisting oflinoleic acid (C18:2 n-6), gamma-linolenic acid (C18:3 n-6),eicosadienoic acid (C20:2 n-6), dihomo-gamma-linolenic acid (C20:3 n-6)(“DGLA”), arachidonic acid (C20:4 n-6) (“AA”), and docosapentaenoic acid(C22:5 n-6, also known as osbond acid).

In particular embodiments, the PUFA free acid solvent system compriseslinoleic acid (C18:2 n-6), gamma-linolenic acid (C18:3 n-6),eicosadienoic acid (C20:2 n-6), dihomo-gamma-linolenic acid (C20:3 n-6)(“DGLA”), arachidonic acid (C20:4 n-6) (“AA”), and docosapentaenoic acid(C22:5 n-6), each present substantially in the free acid form.

In various embodiments, AA, substantially in free acid form, is presentin an amount of no more than about 5% (a/a) of the fatty acids in thePUFA free acid solvent system. In certain embodiments, AA, substantiallyin free acid form, comprises no more than about 4.5% (a/a) of the fattyacids in the PUFA free acid solvent system. In particular embodiments,AA, substantially in free acid form, is present in an amount of no morethan about 4% (a/a) of the fatty acids in the PUFA free acid solventsystem.

In certain embodiments, AA, substantially in the form of the free acid,is present in an amount of no more than about 5% (m/m) of the fattyacids in the PUFA free acid solvent system. In certain embodiments, AAsubstantially in free acid form comprises no more than about 4.5% (m/m)of the fatty acids in the PUFA free acid solvent system. In particularembodiments, AA is present substantially in free acid form in an amountof no more than about 4% (m/m) of the fatty acids in the PUFA free acidsolvent system.

In certain embodiments, total omega-6 polyunsaturated fattyacids—defined as the sum of linoleic acid (C18:2 n-6), gamma-linolenicacid (C18:3 n-6), eicosadienoic acid (C20:2 n-6), dihomo-gamma-linolenicacid (C20:3 n-6), arachidonic acid (C20:4 n-6) and docosapentaenoic acid(C22:5 n-6), each present substantially in the free acid form—compriseno more than about 10% (a/a) of the fatty acids in the PUFA free acidsolvent system. In certain embodiments, total omega-6 polyunsaturatedfatty acids comprise no more than about 10% (m/m) of the fatty acids inthe PUFA free acid solvent system.

In specific embodiments, the PUFA free acid solvent system is given byTable 1, with each species of PUFA identified therein presentsubstantially in free acid form and within the range of about −3 SD toabout +3 SD of the respectively recited average. In certain embodiments,each species of PUFA identified therein is present substantially in freeacid form and within the range of about −2 SD to about +2 SD of therespectively recited average. In certain embodiments, each species ofPUFA identified therein is present substantially in free acid form andwithin the range of about −1 SD to about +1 SD of the respectivelyrecited average. In selected embodiments, each species of PUFAidentified therein is present substantially in free acid form and in anamount approximating the average amount respectively recited in Table 1.

In certain embodiments, the PUFA free acid solvent system is given byTable 2, with each species of PUFA identified therein presentsubstantially in free acid form and within the range of about −3 SD toabout +3 SD of the respectively recited average. In certain embodiments,each species is present substantially in free acid form and within therange of about −2 SD to about +2 SD of the respective average recited inTable 2. In certain embodiments, each PUFA species is presentsubstantially in free acid form and within the range of about −1 SD toabout +1 SD of the respectively recited average. In selectedembodiments, each species of PUFA identified therein is presentsubstantially in free acid form and in an amount approximating theaverage amount respectively recited in Table 2.

In certain embodiments, polyunsaturated fatty acids other than omega-3and omega-6 polyunsaturated fatty acids are present in an amount of nomore than about 5% (a/a). In various embodiments, polyunsaturated fattyacids other than omega-3 and omega-6 polyunsaturated fatty acids arepresent in an amount of no more than about 5% (m/m).

In a variety of embodiments, at least about 90% of each species ofomega-3 PUFA present in the PUFA free acid solvent system is present inthe free acid form. In certain embodiments, at least 91%, at least 92%,at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, even at least 99% of each species of omega-3 PUFA present inthe PUFA free acid solvent system is present in the free acid form. Inexemplary embodiments, at least 90% of the total omega-3 polyunsaturatedfatty acid content in the PUFA free acid solvent system is present inthe free acid form. In certain embodiments, at least 91%, at least 92%,at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, atleast 98%, even at least 99% of the total omega-3 polyunsaturated fattyacid content in the PUFA free acid solvent system is present in the freeacid form.

In various embodiments, at least 90% of each species of omega-6 PUFApresent in the PUFA free acid solvent system is present in the free acidform. In certain embodiments, at least 91%, at least 92%, at least 93%,at least 94%, at least 95%, at least 96%, at least 97%, at least 98%,even at least 99% of each species of omega-6 PUFA in the PUFA free acidsolvent system is present in the free acid form. In exemplaryembodiments, at least 90% of the total omega-6 polyunsaturated fattyacid content in the PUFA free acid solvent system is present in the freeacid form.

In various embodiments, at least 90% of the total polyunsaturated fattyacid content in the PUFA free acid solvent system is present in the freeacid form. In certain embodiments, at least 91%, at least 92%, at least93%, at least 94%, at least 95%, at least 96%, at least 97%, at least98%, even at least 99% of the total polyunsaturated fatty acid in thePUFA free acid solvent system is present in the free acid form.

The PUFA free acid solvent system comprises, in typical embodiments, nomore than about 3% (a/a) saturated fatty acids and no more than about 5%(a/a) mono-unsaturated fatty acids. In various embodiments, the PUFAfree acid solvent system comprises no more than about 3% (m/m) saturatedfatty acids and no more than about 5% (m/m) mono-unsaturated fattyacids.

In typical embodiments, the PUFA free acid solvent system usefullyfurther comprises an antioxidant. In certain embodiments, theantioxidant is butylated hydroxyanisole (BHA). In some embodiments, theantioxidant is alpha-tocopherol. In some embodiments, alpha-tocopherolis present in an amount of about 0.20-about 0.40% (m/m). In variousembodiments, alpha-tocopherol is present in an amount of about0.25-about 0.35% (m/m). In particular embodiments, alpha-tocopherol ispresent in an amount of about 0.27-about 0.33% (m/m). In typicalembodiments, the PUFA free acid solvent system comprises no more thanabout 0.1 ppm ethyl carbamate. In some embodiments, the PUFA free acidsolvent system comprises no more than 0.1 ppm ethyl carbamate. Invarious embodiments, the PUFA free acid solvent system comprises lessthan 0.1 ppm ethyl carbamate.

TABLE 1 PUFA free acid solvent system 10 batch statistics IdentityCommon name AVG SD −3SD −2SD −1SD +1SD +2SD +3SD 1SD Δ 2SD Δ 3SD ΔC18:2(n-6) Linoleic acid 0.61 0.09 0.34 0.43 0.52 0.69 0.78 0.87 0.180.35 0.53 C18:3(n-6) Gamma-linolenic acid 0.15 0.03 0.07 0.10 0.13 0.180.21 0.24 0.06 0.11 0.17 C18:3(n-3) α-Linolenic acid 0.43 0.06 0.23 0.300.36 0.49 0.56 0.62 0.13 0.26 0.39 C18:4(n-3) Moroctic acid 1.56 0.250.81 1.06 1.31 1.81 2.06 2.31 0.50 1.00 1.50 C20:2(n-6) Eicosadienoicacid 0.13 0.05 −0.03 0.02 0.07 0.18 0.23 0.29 0.11 0.21 0.32 C20:3(n-6)Dihomo-gamma- 0.44 0.06 0.28 0.33 0.39 0.50 0.56 0.61 0.11 0.22 0.33linolenic acid C20:4(n-6) Arachidonic acid 3.14 0.58 1.41 1.99 2.57 3.724.29 4.87 1.15 2.30 3.46 C20:3(n-3) Eicosatrienoic acid 0.20 0.04 0.080.12 0.16 0.24 0.28 0.32 0.08 0.16 0.24 C20:4(n-3) Eicosatetraenoic acid2.19 0.24 1.46 1.71 1.95 2.43 2.68 2.92 0.49 0.97 1.46 C20:5(n-3)Eicosapentaenoic acid 56.74 0.56 55.07 55.63 56.19 57.30 57.86 58.421.12 2.23 3.34 (EPA) C21:5(n-3) Heneicosapentaenoic 2.61 0.25 1.85 2.112.36 2.86 3.12 3.37 0.51 1.01 1.52 acid C22:5(n-6) Docosapentaenoic acid0.57 0.21 −0.05 0.16 0.36 0.78 0.98 1.19 0.41 0.83 1.24 C22:5(n-3)Docosapentaenoic acid 5.31 1.06 2.13 3.19 4.25 6.37 7.42 8.48 2.12 4.236.35 (DPA) C22:6(n-3) Docosahexaenoic acid 19.93 0.75 17.68 18.43 19.1820.68 21.43 22.18 1.50 2.99 4.49 (DHA)

TABLE 2 PUFA free acid solvent system 21 batch statistics AVG IdentityCommon name % (a/a) SD −3SD −2SD −1SD +1SD +2SD +3SD 1SD Δ 2SD Δ 3SD ΔC18:2(n-6) Linoleic acid 0.74 0.16 0.26 0.42 0.58 0.90 1.07 1.23 0.320.65 0.97 C18:3(n-6) Gamma-linolenic 0.24 0.11 −0.09 0.02 0.13 0.35 0.460.58 0.22 0.44 0.66 acid C18:3(n-3) α-Linolenic acid 0.54 0.15 0.09 0.240.39 0.69 0.84 0.99 0.30 0.60 0.90 C18:4(n-3) Stearidonic 2.83 1.49−1.63 −0.15 1.34 4.31 5.80 7.28 2.97 5.94 8.92 (moroctic) acidC20:2(n-6) Eicosadienoic acid 0.15 0.04 0.02 0.07 0.11 0.20 0.24 0.280.09 0.17 0.26 C20:3(n-6) Dihomo-gamma- 0.40 0.07 0.18 0.25 0.32 0.470.55 0.62 0.15 0.30 0.45 linolenic acid C20:4(n-6) Arachidonic acid 3.170.51 1.65 2.16 2.67 3.68 4.19 4.70 1.01 2.03 3.04 C20:3(n-3)Eicosatrienoic acid 0.16 0.05 0.01 0.06 0.11 0.21 0.26 0.31 0.10 0.200.31 C20:4(n-3) Eicosatetraenoic 2.13 0.41 0.92 1.32 1.73 2.54 2.94 3.350.81 1.62 2.43 acid C20:5(n-3) Timnodonic acid 55.40 2.13 49.00 51.1353.27 57.53 59.66 61.80 4.26 8.53 12.79 (EPA) C21:5(n-3)Heneicosapentaenoic 2.33 0.34 1.29 1.64 1.98 2.67 3.02 3.36 0.69 1.382.07 acid C22:5(n-6) Docosapentaenoic 0.58 0.16 0.11 0.27 0.43 0.74 0.901.06 0.31 0.63 0.94 acid C22:5(n-3) Docosapentaenoic 4.44 1.16 0.98 2.133.29 5.60 6.75 7.91 2.31 4.62 6.93 acid (DPA) C22:6(n-3) Cervonic acid19.35 1.69 14.28 15.97 17.66 21.04 22.73 24.42 3.38 6.76 10.14 (DHA)

5.2.2. Statins

In various embodiments, at least one statin is present in thepharmaceutical composition in an amount, per ml or per g of PUFA freeacid solvent system, of at least about 2 mg, at least about 2.5 mg, atleast about 5 mg, at least about 10 mg, at least about 15 mg, at leastabout 20 mg, at least about 25 mg, at least about 30 mg, at least about40 mg, at least about 50 mg, at least about 60 mg, at least about 70 mg,at least about 80 mg, at least about 90 mg, even at least about 100 mg,at least about 110 mg, and in certain embodiment, at least about 120 mg.

In selected embodiments, at least one statin is present in thepharmaceutical composition in an amount, per ml or per g of PUFA freeacid solvent system, of from about 2 mg to about 80 mg, from about 5 mgto about 60 mg, or from about 10 mg to about 40 mg, or from about 20 mgto about 30 mg.

In various embodiments, the at least one statin is selected from thegroup consisting of rosuvastatin, pravastatin, lovastatin, simvastatin,atorvastatin, fluvastatin, and pitavastatin.

In typical embodiments, the at least one statin is dissolved in the PUFAfree acid solvent system in an amount that permits administration of atherapeutic amount of statin in a convenient number of oral unit doses,which number of oral unit doses also suffices to deliver a therapeuticdose of omega-3 PUFAs.

Rosuvastatin is marketed under the name Crestor® by Astra Zeneca,Wilmington, Del. In typical embodiments, rosuvastatin is dissolved inthe PUFA free acid solvent system in an amount that permits dailyadministration, in a convenient number of oral doses, of 5 mg, 10 mg, 20mg, 30 mg, 40 mg, 50 mg, 60 mg, 70 mg, 80 mg, even 90 mg or 100 mg. Inparticular embodiments, rosuvastatin is dissolved in the PUFA free acidsolvent system in an amount, per ml or per g of PUFA solvent system, of2.5 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, even 45mg or 50 mg. In certain embodiments, rosuvastatin is dissolved in thePUFA free acid solvent system in an amount, per ml or per g of PUFAsolvent system, of 1.25 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg, 12.5 mg, 15 mg,17.5 mg, 20 mg, 22.5 mg, even 25 mg.

Atorvastatin, which is marketed under the name Lipitor® by Pfizer, NewYork, N.Y., is hydrophobic and is known as a synthetic statin. In someembodiments, atorvastatin is dissolved in the PUFA free acid solventsystem in an amount that permits daily administration, in a convenientnumber of oral doses, of from 2.5 to 100 mg, from 5 to 80 mg, and from10 to 40 mg.

In particular embodiments, atorvastatin is dissolved in the PUFA freeacid solvent system in an amount, per ml or per g of PUFA solventsystem, of 2.5 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40mg, even 45 mg or 50 mg. In certain embodiments, atorvastatin isdissolved in the PUFA free acid solvent system in an amount, per ml orper g of PUFA solvent system, of 1.25 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg,12.5 mg, 15 mg, 17.5 mg, 20 mg, 22.5 mg, even 25 mg.

Pravastatin, which is marketed as Pravachol® by Bristol-Myers Squibb,Princeton, N.J., is hydrophilic. Administered as monotherapy,pravastatin is best absorbed without food. In some embodiments,pravastatin is dissolved in the PUFA free acid solvent system in anamount that permits daily administration, in a convenient number of oraldoses, of from 2.5 to 80 mg, from 5 to 60 mg, or from 10 to 40 mg.

In particular embodiments, pravastatin is dissolved in the PUFA freeacid solvent system in an amount, per ml or per g of PUFA solventsystem, of 2.5 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40mg, even 45 mg or 50 mg. In certain embodiments, pravastatin isdissolved in the PUFA free acid solvent system in an amount, per ml orper g of PUFA solvent system, of 1.25 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg,12.5 mg, 15 mg, 17.5 mg, 20 mg, 22.5 mg, even 25 mg.

Lovastatin, which is marketed under the name Mevacor® by Merck,Whitehouse Station, N.J., is hydrophobic. In some embodiments,lovastatin is dissolved in the PUFA free acid solvent system in anamount that permits daily administration, in a convenient number of oraldoses, of from 2.5 to 100 mg, from 5 to 80 mg, or from 10 to 40 mg.

In particular embodiments, lovastatin is dissolved in the PUFA free acidsolvent system in an amount, per ml or per g of PUFA solvent system, of2.5 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40 mg, even 45mg or 50 mg. In certain embodiments, lovastatin is dissolved in the PUFAfree acid solvent system in an amount, per ml or per g of PUFA solventsystem, of 1.25 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg, 12.5 mg, 15 mg, 17.5mg, 20 mg, 22.5 mg, even 25 mg.

Simvastatin, which is marketed under the name Zocor® by Merck,Whitehouse Station, N.J., is hydrophobic. In some embodiments,simvastatin is dissolved in the PUFA free acid solvent system in anamount that permits daily administration, in a convenient number of oraldoses, of from 1 to 80 mg per day, from 2 to 60 mg, or from 5 to 40 mgper day.

In particular embodiments, simvastatin is dissolved in the PUFA freeacid solvent system in an amount, per ml or per g of PUFA solventsystem, of 2.5 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40mg, even 45 mg or 50 mg. In certain embodiments, simvastatin isdissolved in the PUFA free acid solvent system in an amount, per ml orper g of PUFA solvent system, of 1.25 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg,12.5 mg, 15 mg, 17.5 mg, 20 mg, 22.5 mg, even 25 mg.

Fluvastatin, which is marketed under the name Lescol® by Novartis, EastHanover, N.J., is hydrophilic and is known as a synthetic statin. Invarious embodiments, fluvastatin is dissolved in the PUFA free acidsolvent system in an amount that permits daily administration, in aconvenient number of oral doses, of from 5 to 160 mg, from 10 to 120 mg,and from 20 to 80 mg.

In particular embodiments, fluvastatin is dissolved in the PUFA freeacid solvent system in an amount, per ml or per g of PUFA solventsystem, of 2.5 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40mg, even 45 mg or 50 mg. In certain embodiments, fluvastatin isdissolved in the PUFA free acid solvent system in an amount, per ml orper g of PUFA solvent system, of 1.25 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg,12.5 mg, 15 mg, 17.5 mg, 20 mg, 22.5 mg, even 25 mg.

Pitavastatin is marketed under the name Livalo® by Kowa PharmaceuticalsAmerica, Inc., Montgomery, Ala. and Lilly USA, Indianapolis, Ind. Thedosage of pitavastatin in the pharmaceutical composition can be from 2.5to 80 mg, from 2.5 to 60 mg, or from 5 to 40 mg.

In particular embodiments, pitavastatin is dissolved in the PUFA freeacid solvent system in an amount, per ml or per g of PUFA solventsystem, of 2.5 mg, 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 30 mg, 35 mg, 40mg, even 45 mg or 50 mg. In certain embodiments, pitavastatin isdissolved in the PUFA free acid solvent system in an amount, per ml orper g of PUFA solvent system, of 1.25 mg, 2.5 mg, 5 mg, 7.5 mg, 10 mg,12.5 mg, 15 mg, 17.5 mg, 20 mg, 22.5 mg, even 25 mg.

In typical embodiments, the pharmaceutical composition comprises astatin which is more soluble in a PUFA free acid solvent system than ina PUFA ethyl ester solvent system. In some embodiments, thepharmaceutical composition comprises a hydrophobic statin which is moresoluble in a PUFA free acid solvent system than in a PUFA ethyl estersolvent system. In some embodiments, the pharmaceutical compositioncomprises a hydrophobic statin which is more soluble in a PUFA free acidsolvent system than in a PUFA ethyl ester solvent system.

5.3. Oral Unit Dosage Forms

In another aspect, the pharmaceutical composition is usefully packagedin unit dosage forms for oral administration.

In particular embodiments, the dosage form is a capsule. In certainembodiments, the dosage form is a hard gelatin capsule. In otherembodiments, the dosage form is a soft gelatin capsule.

In various embodiments, the capsule comprises Type A gelatin. In certainembodiments, the capsule comprises Type B gelatin. In some embodiments,the capsule comprises both Type A and Type B gelatin. Sources ofcollagen for the production of either type A or type B gelatin include,but are not limited to, cows, pigs and fish.

In various embodiments, the capsule is a soft gelatin capsule in whichat least about 1% (w/w) of the gelatin is Type A gelatin. In certainembodiments, at least about 2% (w/w), 3% (w/w), 4%, (w/w), 5% (w/w), 6%(w/w), 7% (w/w), 8% (w/w), 9% (w/w), or at least about 10% (w/w) of thegelatin is Type A gelatin. In selected embodiments, at least about 15%(w/w), 20% (w/w), 25% (w/w), 30% (w/w), 35% (w/w), 40% (w/w), 45% (w/w),even at least about 50% (w/w), 55% (w/w), 60% (w/w), 65% (w/w), 70%(w/w), 75% (w/w), 80% (w/w), 85% (w/w), 90% (w/w), 95% or more of thegelatin is Type A gelatin.

In particular embodiments, the gelatin of the capsule consistsessentially of type A gelatin.

In a selection of embodiments, sufficient Type A gelatin is present suchthat the capsule, when lacking an exterior coating such as thosedescribed below, disintegrates within a time period of not more than 30minutes in purified water at 37° C. after storage for at least 3 monthsat 40° C. In certain embodiments, the capsule is a soft gelatin capsulecomprising sufficient Type. A gelatin such that the capsule, uncoated,disintegrates within a time period of not more than 30 minutes inpurified water at 37° C. after storage for at least 6 months at 40° C.

In various embodiments, the capsule is a soft gelatin capsule comprisingsufficient Type A gelatin such that the capsule, when uncoated,disintegrates within a time period of not more than 30 minutes inpurified water at 37° C. after storage for at least 3 months at 30° C.In certain embodiments, the capsule is a soft gelatin capsule comprisingsufficient Type A gelatin such that the capsule, when not coated,disintegrates within a time period of not more than 30 minutes inpurified water at 37° C. after storage for at least 6 months at 30° C.In some embodiments, the capsule is a soft gelatin capsule comprisingsufficient Type A gelatin such that the capsule in an uncoated statedisintegrates within a time period of not more than 30 minutes inpurified water at 37° C. after storage for at least 9 months at 30° C.In some embodiments, the capsule is a soft gelatin capsule comprisingsufficient Type A gelatin such that the capsule, if not further coated,disintegrates within a time period of not more than 30 minutes inpurified water at 37° C. after storage for at least 12 months at 30° C.

In certain embodiments, the Type A gelatin is porcine Type A gelatin.

In some embodiments, the capsule is a reduced cross-linked gelatincapsule, such as those described in U.S. Pat. No. 7,485,323,incorporated herein by reference in its entirety. In a variety ofembodiments, capsules are made from substances that are not animalby-products, such as alginate, agar-agar, carrageenan, pectin, konjak,guar gum, food starch, modified corn starch, potato starch, and tapioca.Non-animal sources of materials that can be used to make capsules aredescribed in U.S. Patent Publication No. 2011/0117180, incorporatedherein by reference. In some embodiments, Vegicaps® Capsules (Catalent)are used.

In certain embodiments, the capsule comprises a chemically-modifiedgelatin. In various embodiments, the chemically-modified gelatin is asuccinylated gelatin.

In certain capsular oral unit dosage form embodiments, the capsule isuncoated. In a variety of embodiments, the capsule is coated.

In certain coated capsule embodiments, the capsule is coated with acoating on the exterior of the capsule that causes the encapsulatedpharmaceutical composition to be released in a time-dependent manner. Invarious embodiments, release of the pharmaceutical composition isdelayed for at least 15 minutes after ingestion. In particularembodiments, release of the pharmaceutical composition is delayed for atleast 30 minutes after ingestion. In other embodiments, release of thefatty acid composition is delayed for about 30 minutes-about 60 minutesafter ingestion. In various coated embodiments, the coating is selectedfrom cellulose acetate trimellitate, cellulose acetate phthalate andpoly(ethylacrylate-methylacrylate). In some embodiments, the coating isa neutral polyacrylate such as poly(ethylacrylatemethylmethacrylate),such as Eudragit NE 30-D (Evonik Industries AG), which has an averagemolecular weight of about 800,000.

In certain embodiments, capsules are coated as described in U.S. Pat.Nos. 5,792,795 and 5,948,818, the disclosures of which are incorporatedherein by reference. In certain embodiments, the dosage form is a coatedsoft gelatin capsule comprising porcine type A gelatin, as described inU.S. Pat. No. 7,960,370, incorporated herein by reference.

In various embodiments, the oral unit dosage form contains from about100 mg to about 2000 mg of the pharmaceutical composition describedherein. In some embodiments, the oral dosage form contains about 250 mgof the pharmaceutical composition. In some embodiments, the oral dosageform contains about 500 mg of the pharmaceutical composition. In certainembodiments, the oral dosage form contains about 750 mg of thepharmaceutical composition. In some embodiments, the oral dosage formcontains about 1000 mg of the pharmaceutical composition. In otherembodiments, the oral dosage form contains about 1500 mg of thepharmaceutical composition. In certain embodiments, the unit dosage formcontains nonintegral weight amounts of pharmaceutical composition,typically between 100 mg and 2000 mg.

In some embodiments, the dosage form encapsulates PUFAs in an amount ofabout 50 mg to about 5000 mg, about 75 mg to about 2500 mg, or about 100mg to about 1000 mg, for example about 75 mg, about 100 mg, about 125mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about 375mg, about 400 mg, about 425 mg, about 450 mg, about 475 mg, about 500mg, about 525 mg, about 550 mg, about 575 mg, about 600 mg, about 625mg, about 650 mg, about 675 mg, about 700 mg, about 725 mg, about 750mg, about 775 mg, about 800 mg, about 825 mg, about 850 mg, about 875mg, about 900 mg, about 925 mg, about 950 mg, about 975 mg, about 1000mg, about 1025 mg, about 1050 mg, about 1075 mg, about 1100 mg, about1025 mg, about 1050 mg, about 1075 mg, about 1200 mg, about 1225 mg,about 1250 mg, about 1275 mg, about 1300 mg, about 1325 mg, about 1350mg, about 1375 mg, about 1400 mg, about 1425 mg, about 1450 mg, about1475 mg, about 1500 mg, about 1525 mg, about 1550 mg, about 1575 mg,about 1600 mg, about 1625 mg, about 1650 mg, about 1675 mg, about 1700mg, about 1725 mg, about 1750 mg, about 1775 mg, about 1800 mg, about1825 mg, about 1850 mg, about 1875 mg, about 1900 mg, about 1925 mg,about 1950 mg, about 1975 mg, about 2000 mg, about 2025 mg, about 2050mg, about 2075 mg, about 2100 mg, about 2125 mg, about 2150 mg, about2175 mg, about 2200 mg, about 2225 mg, about 2250 mg, about 2275 mg,about 2300 mg, about 2325 mg, about 2350 mg, about 2375 mg, about 2400mg, about 2425 mg, about 2450 mg, about 2475 mg or about 2500 mg.

In various embodiments, the pharmaceutical composition present in theunit dosage form is stable at room temperature (about 23° C. to 27° C.,or about 25° C.) and about 60% relative humidity for a period of atleast six months, at least one year, or at least two years. By “stable”is intended that the solubilized statin does not precipitate out ofsolution to any appreciable degree, for example, in amounts of less than10%, preferably less than 5%, of the originally dissolved statin.

5.4. Dosage Kits

In another aspect, a plurality of unit dosage forms as above-describedmay usefully be packaged together in a dosage kit to increase ease ofuse and patient compliance.

In certain embodiments, the dosage kit is a bottle. In otherembodiments, the plurality of dosage forms is packaged in blister packs,a plurality of which blister packs may optionally be packaged togetherin a box or other enclosure. Typically, whether in a bottle or one ormore blister packs, the plurality of unit dosage forms is sufficient for30 days, 60 days, or 90 days of dosing. Thus, in selected embodiments,in which the unit dosage form is a capsule that encapsulatesapproximately one gram of the pharmaceutical composition as describedherein above, the dosage kit comprises 30, 60, 90, 120, 150, 180, 240,270, or 300 such capsules.

In various embodiments, the plurality of unit dosage forms is packagedunder an inert gas, such as nitrogen or a noble gas, or is packagedunder vacuum.

5.5. Dosing and Administration

In typical embodiments, the pharmaceutical composition is administeredfrom 1 to 4 times a day, e.g. twice a day, with each dose comprisingfrom 1 to 10 unit doses, such as capsules, as described herein. Intypical embodiments, the administration is oral administration, althoughother routes of administration may be used.

In certain embodiments, at least about 2 g of the pharmaceuticalcomposition is administered per day. In some embodiments, at least about3 g of the pharmaceutical composition is administered per day. Incertain embodiments, at least about 4 g of the pharmaceuticalcomposition is administered per day. Typically, the pharmaceutical isadministered as a plurality of unit dosage forms, such as thosedescribed above. Thus, in certain embodiments, at least 2 unit dosageforms, each comprising 1 g of the pharmaceutical composition, areadministered per day. In various embodiments, at least 3 unit dosageforms, each comprising 1 g of the pharmaceutical composition, areadministered per day. In particular embodiments, at least 4 unit dosageforms, each comprising 1 g of the pharmaceutical composition, areadministered per day.

5.6. Methods of Treatment

In another aspect, methods of treating a blood lipid disorder arepresented, comprising administering a pharmaceutical composition orcompositions as described herein, typically in the form of unit dosageforms as described herein, in an amount and for a duration sufficient totreat the blood lipid disorder. Blood lipid disorders include, but arenot limited to hypertriglyceridemia, dyslipidemias,hypercholesterolemia, coronary heart disease (CHD), heart failure,myocardial infarction, cardiac arrhythmias, ischemic dementia,hypertension, coagulation related disorders, nephropathy, retinopathy,vascular disease, atherosclerotic disease and related conditions.Methods of the disclosure are also directed to the treatment and/orprevention and/or reduction of cardiac events, cardiovascular events andsymptoms.

The present combination of a statin and omega-3 fatty acids in a singlecomposition may allow for a greater effect than any expected combined oradditive effect of the two drugs alone. Moreover, the combined oradditive effect of the two drugs may depend on the initial level oflipid parameter in the blood of a subject.

In some embodiments, upon treatment in accordance with the presentinvention, the subject exhibits an improvement in one or more parametersassociated with a blood lipid disorder. Non-limiting examples of suchimproved parameters are reduced triglyceride levels compared to baselineor placebo control; reduced Apo B levels compared to baseline or placebocontrol; reduced Apo CIII levels compared to baseline or placebocontrol; increased HDL-C levels compared to baseline or placebo control;a reduction in LDL-C levels compared to baseline or placebo control; areduction in non-HDL-C levels compared to baseline or placebo control; areduction in vLDL levels compared to baseline or placebo control; anincrease in Apo A-I levels compared to baseline or placebo control; anincrease in Apo A-I/Apo B ratio compared to baseline or placebo control;a reduction in lipoprotein a levels compared to baseline or placebocontrol; a reduction in LDL particle number compared to baseline orplacebo control; a reduction in LDL particle size compared to baselineor placebo control; a reduction in remnant-like particle cholesterolcompared to baseline or placebo control; a reduction in oxidized LDLcompared to baseline or placebo control; a reduction in fasting plasmaglucose (FPG) compared to baseline or placebo control; a reduction inhemoglobin A (HbA) compared to baseline or placebo control; a reductionin homeostasis model insulin resistance compared to baseline or placebocontrol; a reduction in lipoprotein associated phospholipase A2 comparedto baseline or placebo control; a reduction in intracellular adhesionmolecule-1 compared to baseline or placebo control; a reduction ininterleukin-2 compared to baseline or placebo control; a reduction inplasminogen activator inhibitor-1 compared to baseline or placebocontrol; a reduction in high sensitivity C-reactive protein (hsCRP)compared to baseline or placebo control; an increase in serumphospholipid EPA compared to baseline or placebo control; an increase inred blood cell membrane EPA compared to baseline or placebo control; areduction in serum or plasma arachidonic acid (AA) level compared tobaseline or placebo control; an increase in plasma or red blood cellmembrane EPA/arachidonic acid (AA) ratio; and an increase inomega-3:omega-6 ratio.

Measurements of parameters associated with a blood lipid disorder can bemade using methods known in the art. Measurements can be made todetermine baseline levels of one or more parameters associated with ablood lipid disorder, prior to treatment, or during the course oftreatment.

For example, triglycerides, total cholesterol, HDL-C and fasting bloodsugar can be sampled from serum or plasma and analyzed using standardphotometry techniques. VLDL-TG, LDL-C and VLDL-C can be calculated ordetermined using serum lipoprotein fractionation by preparativeultracentrifugation and subsequent quantitative analysis byrefractometry or by analytic ultracentrifugal methodology. Apo A1, Apo Band hsCRP can be determined from serum using standard nephelometrytechniques. Lipoprotein (a) can be determined from serum using standardturbidimetric immunoassay techniques. LDL particle number and particlesize can be determined using nuclear magnetic resonance (NMR)spectrometry. Remnants lipoproteins and LDL-phospholipase A2 can bedetermined from EDTA plasma or serum and serum, respectively, usingenzymatic immunoseparation techniques. Oxidized LDL, intercellularadhesion molecule-1 and interleukin-2 levels can be determined fromserum using standard enzyme immunoassay techniques. These techniques aredescribed in detail in standard textbooks, for example TietzFundamentals of Clinical Chemistry, 6th Ed. (Burtis, Ashwood and BorterEds.), WB Saunders Company.

In some embodiments, a subject fasts for up to 12 hours prior to bloodsample collection, for example about 10 hours.

5.6.1. Treatment of Severe Hypertriglyceridemia (≧500 mg/dL)

In various embodiments, the method is a method of treating severe (≧500mg/dL) hypertriglyceridemia, the method comprising orally administeringthe pharmaceutical composition described herein to a patient havingpre-treatment serum or plasma triglyceride levels≧500 mg/dL, in anamount and for a duration effective to reduce serum or plasmatriglycerides below pre-treatment levels.

In certain embodiments, the method comprises administering thepharmaceutical composition in an amount and for a duration effective toreduce serum or plasma triglyceride levels by at least about 5%, 6%, 7%,8%, or at least about 9% below pre-treatment levels. In certainembodiments, the composition is administered in an amount and for aduration effective to reduce serum or plasma triglyceride levels by atleast 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18% or 19% belowpre-treatment levels. In particular embodiments, the composition isadministered in an amount and for a duration effective to reduce serumor plasma triglyceride levels by at least about 20% below pre-treatmentlevels. In various embodiments, the pharmaceutical composition isadministered in an amount and for a duration effective to reduce serumor plasma triglycerides by at least about 25%, 30%, 35%, 40%, 45%, evenat least about 50% below pre-treatment levels.

In a series of embodiments, the pharmaceutical composition isadministered in an amount and for a duration effective to reduce serumor plasma triglyceride levels by at least about 50 mg/dL, 60 mg/dL, 70mg/dL, 80 mg/dL, 90 mg/dL, even at least about 100 mg/dL. In certainembodiments, the composition is administered in an amount and for aduration effective to reduce serum or plasma triglyceride levels by atleast about 110 mg/dL, 120 mg/dL, 130 mg/dL, 140 mg/dL, even at leastabout 150 mg/dL. In specific embodiments, the pharmaceutical compositionis administered in an amount and for a duration effective to reduceserum or plasma triglyceride levels by at least about 160 mg/dL, 170mg/dL, 180 mg/dL, even at least about 190 mg/dL or 200 mg/dL.

In some embodiments, the pharmaceutical composition described herein isadministered in an amount and for a duration sufficient to reducenon-HDL-cholesterol by at least about 1%, at least about 2%, at leastabout 3%, 4%, 5%, even at least about 7%, 8%, 9%, or at least about 10%below pre-treatment levels. In certain embodiments, the pharmaceuticalcomposition is administered in an amount and for a duration sufficientto reduce non-HDL-cholesterol by at least about 15%, at least about 20%,at least about 25%, at least about 30%, at least about 35%, at leastabout 40% below pre-treatment levels.

In various embodiments, the pharmaceutical composition is administeredin an amount and for a duration effective to increase HDL-c levels by atleast about 1% above pre-treatment levels. In certain embodiments, thepharmaceutical composition is administered in an amount and for aduration sufficient to increase HDL-c by at least about 2%, 3%, 4%, evenat least about 5%, 6%, 7%, 8%, 9%, or 10% above pre-treatment levels.

In certain embodiments, the pharmaceutical composition is administeredin an amount and for a duration effective to reduce the totalcholesterol:HDL-c (“TC/HDL”) ratio by at least about 1% belowpre-treatment levels. In some embodiments, the pharmaceuticalcomposition is administered in an amount and for a duration sufficientto reduce the TC/HDL ratio by at least about 2%, 3%, 4%, 5%, 6%, 7%, 8%,even at least about 9% or at least about 10% below pre-treatment levels.

In some embodiments, the pharmaceutical composition is administered inan amount and for a duration effective to decrease VLDL-c levels by atleast about 5%, 6%, 7%, 8%, 9%, or at least about 10% belowpre-treatment levels. In certain embodiments, the pharmaceuticalcomposition is administered in an amount and for a duration sufficientto decrease VLDL-c levels by at least about 11%, 12%, 13%, 14%, 15%,16%, 17%, even at least about 18%, 19%, or 20% below pre-treatmentlevels. In particular embodiments, the pharmaceutical composition isadministered in an amount and for a duration sufficient to decreaseVLDL-c levels by at least about 21%, 22%, 23%, 24%, even at least about25% below pre-treatment levels.

In a variety of embodiments, the pharmaceutical composition isadministered in an amount and for a duration effective to decreaseApoCIII levels. In certain embodiments, the pharmaceutical compositionis administered in an amount and for a duration sufficient to decreaseApoCIII levels by at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, even atleast about 8%, 9% or 10% below pre-treatment levels.

In some embodiments, the pharmaceutical composition is administered inan amount and for a duration effective to increase plasma EPA levels byat least 100% above pre-treatment levels. In certain embodiments, thepharmaceutical composition is administered in an amount and for aduration effective to increase plasma EPA levels by at least about 200%,250%, 300%, even at least about 350%, 400%, 450% or at least about 500%above pre-treatment levels. In selected embodiments, the pharmaceuticalcomposition is administered for a time and in an amount effective toincrease plasma EPA levels by at least about 550%, 600%, 650%, even atleast about 700% above pre-treatment levels.

In various embodiments, the pharmaceutical composition is administeredin an amount and for a duration effective to increase plasma DHA levelsby at least about 50% above pre-treatment levels. In particularembodiments, the pharmaceutical composition is administered in an amountand for a duration effective to increase plasma DHA levels by at leastabout 55%, 60%, 65%, 70%, even at least about 75%, 80%, 85%, or 90%above pre-treatment levels.

In a series of embodiments, the pharmaceutical composition isadministered in an amount and for a duration effective to increaseplasma DPA levels by at least about 50% above pre-treatment levels. Insome embodiments, the pharmaceutical composition is administered in anamount and for a duration effective to increase plasma DPA levels by atleast about 55%, 60%, 65%, 70%, 75%, even at least about 80%, 85%, 90%,95%, or 100% above pre-treatment levels. In selected embodiments, thepharmaceutical composition is administered in an amount and for aduration effective to increase plasma DPA levels by at least about 110%,120%, even at least about 125% above pre-treatment levels.

In a series of embodiments, the pharmaceutical composition isadministered in an amount and for a duration effective to reducearachidonic acid (AA) concentration in plasma by at least about 5% belowpre-treatment levels. In certain embodiments, the pharmaceuticalcomposition is administered in an amount and for a duration effective toreduce arachidonic (AA) concentration in plasma by at least about 6%,7%, 8%, 9%, 10%, even at least about 11%, 12%, 13%, 14%, even at leastabout 15%, 16%, 17%, 18%, 19%, 20%, or 21%, 22%, 23%, 24% even at leastabout 25% below pre-treatment levels.

In certain embodiments, the pharmaceutical composition is administeredin an amount, and for a duration, effect to reduce plasma arachidonicacid concentration by at least about 25 μg/mL. In some embodiments, thepharmaceutical composition is administered in an amount and for aduration sufficient to reduce plasma AA levels by at least about 50μg/mL, 55 μg/mL, 60 μg/mL, 65 μg/mL, even at least about 70 μg/mL, 75μg/mL, 80 μg/mL, 85 μg/mL, 90 μg/mL, even at least about 95 μg/mL or 100μg/mL.

In certain embodiments, the effective amount is at least about 2 g perday. In various embodiments, the effective amount is at least about 3 gper day. In particular embodiments, the effective amount is at leastabout 4 g per day. In typical embodiments, the effective amount is about2 g per day. In certain embodiments, the effective amount is about 4 gper day.

In typical embodiments, the pharmaceutical composition is administeredfor at least 30 days. In certain embodiments, the pharmaceuticalcomposition is administered for at least 60 days. In particularembodiments, the pharmaceutical composition is administered for at least90 days, 120 days, 180 days, 240 days, or at least 360 days. In certainembodiments, the pharmaceutical composition is administeredindefinitely.

In some embodiments, the pharmaceutical composition is administereddaily. In other embodiments, the pharmaceutical composition isadministered every other day.

In particular embodiments, the daily dosage of pharmaceuticalcomposition is administered in a single daily dose. In otherembodiments, the pharmaceutical composition is administered in divideddoses, with the daily dose divided into two administrations, threeadministrations, or even four administrations, over the course of theday.

In certain embodiments, the pharmaceutical composition is administeredwith food. In certain embodiments, the pharmaceutical composition isadministered with a low fat meal. In other embodiments, thepharmaceutical composition is administered without food. In certainembodiments, the pharmaceutical composition is administered in thefasting state.

5.6.2. Treatment of Hypertriglyceridemia (200-500 Mg/dL)

In another series of treatment embodiments, methods are provided fortreating patients who have pre-treatment serum or plasma triglyceridelevels of about 200 mg/dL to about 500 mg/dL. In certain embodiments,the patients are already on statin therapy; in these patients, thepre-treatment serum or plasma triglyceride levels are those measuredduring statin treatment, prior to administration of the pharmaceuticalcompositions described herein.

The method comprises orally administering the pharmaceutical compositiondescribed herein to a patient having pre-treatment serum or plasmatriglyceride levels of about 200 mg/dL to about 500 mg/dL, in an amountand for a duration effective to reduce serum or plasma triglyceridesbelow pre-treatment levels.

In certain embodiments, the pharmaceutical composition described hereinis administered in an amount and for a duration sufficient to reduceserum or plasma triglyceride levels by at least about 5% belowpre-treatment levels. In various embodiments, the pharmaceuticalcomposition is administered in an amount and for a duration sufficientto reduce serum or plasma triglyceride levels by at least about 6%, 7%,8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, even at least about 16%, 17%, 18%,19%, or at least about 20% below pre-treatment levels.

In some embodiments, the pharmaceutical composition described herein isadministered in an amount and for a duration sufficient to reducenon-HDL-cholesterol by at least about 1%, at least about 2%, at leastabout 3%, 4%, 5%, even at least about 7%, 8%, 9%, or at least about 10%below pre-treatment levels. In certain embodiments, the pharmaceuticalcomposition is administered in an amount and for a duration sufficientto reduce non-HDL-cholesterol by at least about 15%, at least about 20%,at least about 25%, at least about 30%, at least about 35%, at leastabout 40% below pre-treatment levels.

In a series of embodiments, the pharmaceutical composition isadministered in an amount and for a duration sufficient to raise HDL-clevels by at least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, even at least about8%, 9%, or 10% or more above pre-treatment levels.

In some embodiments, the pharmaceutical composition is administered inan amount and for a duration effective to increase plasma EPA levels byat least 100% above pre-treatment levels. In certain embodiments, thepharmaceutical composition is administered in an amount and for aduration effective to increase plasma EPA levels by at least about 200%,250%, 300%, even at least about 350%, 400%, 450% or at least about 500%above pre-treatment levels. In selected embodiments, the pharmaceuticalcomposition is administered for a time and in an amount effective toincrease plasma EPA levels by at least about 550%, 600%, 650%, even atleast about 700% above pre-treatment levels.

In various embodiments, the pharmaceutical composition is administeredin an amount and for a duration effective to increase plasma DHA levelsby at least about 50% above pre-treatment levels. In particularembodiments, the pharmaceutical composition is administered in an amountand for a duration effective to increase plasma DHA levels by at leastabout 55%, 60%, 65%, 70%, even at least about 75%, 80%, 85%, or 90%above pre-treatment levels.

In a series of embodiments, the pharmaceutical composition isadministered in an amount and for a duration effective to increaseplasma DPA levels by at least about 50% above pre-treatment levels. Insome embodiments, the pharmaceutical composition is administered in anamount and for a duration effective to increase plasma DPA levels by atleast about 55%, 60%, 65%, 70%, 75%, even at least about 80%, 85%, 90%,95%, or 100% above pre-treatment levels. In selected embodiments, thepharmaceutical composition is administered in an amount and for aduration effective to increase plasma DPA levels by at least about 110%,120%, even at least about 125% above pre-treatment levels.

In a series of embodiments, the pharmaceutical composition isadministered in an amount and for a duration effective to reducearachidonic acid (AA) concentration in plasma by at least about 5% belowpre-treatment levels. In certain embodiments, the pharmaceuticalcomposition is administered in an amount and for a duration effective toreduce arachidonic (AA) concentration in plasma by at least about 6%,7%, 8%, 9%, 10%, even at least about 11%, 12%, 13%, 14%, even at leastabout 15%, 16%, 17%, 18%, 19%, 20%, or 21%, 22%, 23%, 24% even at leastabout 25% below pre-treatment levels.

In certain embodiments, the pharmaceutical composition is administeredin an amount, and for a duration, effect to reduce plasma arachidonicacid concentration by at least about 25 μg/mL. In some embodiments, thepharmaceutical composition is administered in an amount and for aduration sufficient to reduce plasma AA levels by at least about 50μg/mL, 55 μg/mL, 60 μg/mL, 65 μg/mL, even at least about 70 μg/mL, 75μg/mL, 80 μg/mL, 85 μg/mL, 90 μg/mL, even at least about 95 μg/mL or 100μg/mL.

In various embodiments, the pharmaceutical composition described inSection 5.2 herein is administered in unit dosage forms as described inSection 5.3 above.

In various embodiments, the pharmaceutical composition is administeredin an amount of at least about 1 g per day. In some embodiments, thepharmaceutical composition is administered in an amount of at leastabout 2 g/day. In certain embodiments, the pharmaceutical composition isadministered in an amount of at least about 3 g/day. In particularembodiments, the pharmaceutical composition is administered in an amountof at least about 4 g/day. In typical embodiments, the pharmaceuticalcomposition is administered in an amount of about 2 g/day. In certainembodiments, the pharmaceutical composition is administered in an amountof about 3 g/day or about 4 g per day.

5.6.3. Treatment of Primary Hyperlipidemia or Mixed Dyslipidemia

In a series of treatment embodiments, methods are provided for treatingpatients who have primary hyperlipidemia or mixed dyslipidemia. Themethod comprises orally administering the pharmaceutical compositiondescribed herein to a patient having primary hyperlipidemia or mixeddyslipidemia, in an amount and for a duration effective to reduce one ormore of elevated total-cholesterol, elevated LDL-C, elevated ApoB,non-HDL-C, and elevated triglyceride levels, and/or to increase HDL-C.

In various embodiments, the pharmaceutical composition is administeredin an amount of at least about 1 g per day. In some embodiments, thepharmaceutical composition is administered in an amount of at leastabout 2 g/day. In certain embodiments, the pharmaceutical composition isadministered in an amount of at least about 3 g/day. In particularembodiments, the pharmaceutical composition is administered in an amountof at least about 4 g/day. In typical embodiments, the pharmaceuticalcomposition is administered in an amount of about 2 g/day. In certainembodiments, the pharmaceutical composition is administered in an amountof about 3 g/day or about 4 g per day.

5.6.4. Treatment to Increase Plasma EPA:AA Ratios

Methods are also provided for increasing the EPA:AA ratio. The methodscomprise administering the pharmaceutical composition described inSection 5.2 herein to a patient having an EPA:AA ratio below about 0.25,in an amount and for duration sufficient to increase the patient'sEPA:AA ratio to at least about 0.25. In some embodiments, thepharmaceutical composition is administered in an amount and for aduration sufficient to increase the patient's EPA:AA ratio to at leastabout 0.3, at least about 0.35, at least about 0.40, at least about0.45, at least about 0.50, even to a level of at least about 0.55, 0.60,0.61, 0.62, 0.63, 0.64, or 0.65.

In certain embodiments, the method comprises administering thepharmaceutical composition in an amount and for a duration effective toincrease plasma EPA levels by at least 100% above pre-treatment levels.In certain embodiments, the pharmaceutical composition is administeredin an amount and for a duration effective to increase plasma EPA levelsby at least about 200%, 250%, 300%, even at least about 350%, 400%, 450%or at least about 500% above pre-treatment levels. In selectedembodiments, the pharmaceutical composition is administered for a timeand in an amount effective to increase plasma EPA levels by at leastabout 550%, 600%, 650%, even at least about 700% above pre-treatmentlevels.

In various embodiments, the pharmaceutical composition is administeredin an amount and for a duration effective to increase plasma DHA levelsby at least about 50% above pre-treatment levels. In particularembodiments, the pharmaceutical composition is administered in an amountand for a duration effective to increase plasma DHA levels by at leastabout 55%, 60%, 65%, 70%, even at least about 75%, 80%, 85%, or 90%above pre-treatment levels.

In a series of embodiments, the pharmaceutical composition isadministered in an amount and for a duration effective to reducearachidonic acid (AA) concentration in plasma by at least about 5% belowpre-treatment levels. In certain embodiments, the pharmaceuticalcomposition is administered in an amount and for a duration effective toreduce arachidonic (AA) concentration in plasma by at least about 6%,7%, 8%, 9%, 10%, even at least about 11%, 12%, 13%, 14%, even at leastabout 15%, 16%, 17%, 18%, 19%, 20%, or 21%, 22%, 23%, 24% even at leastabout 25% below pre-treatment levels.

In certain embodiments, the pharmaceutical composition is administeredin an amount, and for a duration, effect to reduce plasma arachidonicacid concentration by at least about 25 μg/mL. In some embodiments, thepharmaceutical composition is administered in an amount and for aduration sufficient to reduce plasma AA levels by at least about 50μg/mL, 55 μg/mL, 60 μg/mL, 65 μg/mL, even at least about 70 μg/mL, 75μg/mL, 80 μg/mL, 85 μg/mL, 90 μg/mL, even at least about 95 μg/mL or 100μg/mL.

In various embodiments, the pharmaceutical composition described inSection 5.2 herein is administered in unit dosage forms as described inSection 5.3 above.

In various embodiments, the pharmaceutical composition is administeredin an amount of at least about 1 g per day. In some embodiments, thepharmaceutical composition is administered in an amount of at leastabout 2 g/day. In certain embodiments, the pharmaceutical composition isadministered in an amount of at least about 3 g/day. In particularembodiments, the pharmaceutical composition is administered in an amountof at least about 4 g/day. In typical embodiments, the pharmaceuticalcomposition is administered in an amount of about 2 g/day. In certainembodiments, the pharmaceutical composition is administered in an amountof about 3 g/day or about 4 g per day.

5.6.5. Treatment to Lower Serum or Plasma ApoCIII Levels

Methods are also provided for increasing a patient's serum or plasmaApoCIII levels, without regard to the patient's pre-treatment plasmatriglyceride levels. The methods comprise administering thepharmaceutical composition described in Section 5.2 herein to a patientin need of lower ApoCIII levels, in an amount and for durationsufficient to decrease the patient's serum or plasma ApoCIII levels. Intypical embodiments, the patient is at risk for cardiovascular heartdisease.

In certain embodiments, the pharmaceutical composition is administeredin an amount and for a duration sufficient to decrease ApoCIII levels byat least about 1%, 2%, 3%, 4%, 5%, 6%, 7%, even at least about 8%, 9% or10% below pre-treatment levels.

5.6.6. Other Methods of Treatment

In another aspect, the pharmaceutical compositions described herein isused to treat other disorders, including one or more of nonalcoholicsteatohepatitis (NASH), hyperlipoproteinemia, including type IIIhyperlipoproteinemia, and metabolic syndrome.

In certain embodiments, the pharmaceutical composition is used to reduceresistance to platelet aggregation inhibitors, such as Plavix, includinguse in the methods described in U.S. patent application Ser. No.13/620,312, the disclosure of which is incorporated herein by referencein its entirety.

6. EXAMPLES Example 1 Solubility of Statins in Various pH Buffers

50 mg of atorvastatin was added to 5 mL of water, 0.1N NaCl, 0.1N NaOH,0.1N HCl (pH 1.2), acid phthalate buffer (pH 2.2), acid phthalate buffer(pH 4.0), neutralized phthalate buffer (pH 5.0), phosphate buffer (pH7.0), or alkaline borate buffer (pH 9.0). The solutions were sonicatedfor 10 minutes and allowed to sit overnight. Aliquots of the samplesolutions were centrifuged and an aliquot of the supernatant from eachsample was assayed for atorvastatin solubility. Samples were thenallowed to sit for 1 week when they were again assayed to determineatorvastatin solubility. The results of solubility measurements aredisplayed in Table 3.

50 mg of rosuvastatin was added to 5 mL of water, 0.1N NaCl, 0.1N NaOH,0.1N HCl (pH 1.2), acid phthalate buffer (pH 2.2), acid phthalate buffer(pH 4.0), neutralized phthalate buffer (pH 5.0), phosphate buffer (pH7.0), or alkaline borate buffer (pH 9.0). The solutions were sonicatedfor 10 minutes and allowed to sit overnight. Aliquots of the samplesolutions were centrifuged and an aliquot of the supernatant from eachsample was assayed for rosuvastatin solubility. Samples were thenallowed to sit for 1 week when they were again assayed to determinerosuvastatin solubility. The results of solubility measurements aredisplayed in Table 3.

TABLE 3 Atorvastatin Calcium and Rosuvastatin Calcium Solubility DataAtorvastatin Rosuvastatin Solution T = 0 T = 1 Week T = 0 T = 1 WeekWater 0.146 0.138 7.08 8.822 0.1N HCl 0.019 0.012 0.223 0.104 pH 2.20.041 0.015 0.773 9.343 pH 4.0 0.074 0.06 1.901 0.002 pH 5.0 0.115 0.14.188 4.790 pH 7.0 0.286 0.285 7.963 10.654 pH 9.0 0.247 0.225 4.5714.712 0.1N NaOH 0.246 0.352 8.058 0.018

The results showed that Rosuvastatin displayed significant aqueoussolubility at time zero, with the maximal solubility at about pH 7.0.Atorvastatin showed a lower overall aqueous solubility thanrosuvastatin, but the solubility also displayed a maximum at pH 7.0.

Example 2 Solubility of Statins in PUFA Free Acid Solvent System

Samples of 20-30 mg of atorvastatin and 20-30 mg of rosuvastatin wereeach dissolved in 2 g of a PUFA free acid solvent system. Thecomposition of the solvent system is given in Table 4, below (batch#37225).

TABLE 4 PUFA free acid solvent system (batch # 37225) Identity Commonname area % C18:2(n-6) Linoleic acid 0.59 C18:3(n-6) Gamma-linolenicacid 0.12 C18:3(n-3) α-Linolenic acid 0.38 C18:4(n-3) Moroctic acid 1.16C20:2(n-6) Eicosadienoic acid 0.12 C20:3(n-6) Dihomo-gamma-linolenicacid 0.45 C20:4(n-6) Arachidonic acid 2.84 C20:3(n-3) Eicosatrienoicacid 0.22 C20:4(n-3) Eicosatetraenoic acid 2.11 C20:5(n-3) EPA 55.81C21:5(n-3) Heneicosapentaenoic acid 2.72 C22:5(n-6) Docosapentaenoicacid 0.72 C22:5(n-3) DPA 5.46 C22:6(n-3) DHA 19.45

Initial solubility was measured, and results of solubility measurementare displayed in Table 5.

TABLE 5 Initial solubility in PUFA free acid solvent systemConcentration (mg/mL) Solution Atorvastatin Rosuvastatin PUFA free acidsolvent system 7.85 12.70 Batch #37225

Example 3 Solubility of Statins in Omega-3 Ethyl Ester Solvent System

20 mg of atorvastatin and 20 mg of rosuvastatin were each added to 2 gof fill extracted from capsules of Lovaza, in which the PUFA species arepresent substantially in the ethyl ester form. The samples were thensonicated for 1 hour and incubated overnight, after which they displayedexcess un-dissolved solids. The samples were centrifuged and thesupernatant assayed by HPLC for the respective statin. The results ofthe solubility measurement are displayed in Table 6. As is apparent fromcomparing Tables 5 and Table 6, atorvastatin and rosuvastatin weresolubilized to a significantly greater extent in a PUFA free acidsolvent system than in a PUFA ethyl ester solvent system.

TABLE 6 Initial solubility in PUFA ethyl ester solvent systemConcentration (mg/mL) Solution Atorvastatin Rosuvastatin Lovaza 0.0050.001

Example 4 Comparison of Solubility and Stability of Statins Dissolved inOmega-3 Free Fatty Acids and Omega-3 Ethyl Esters

Solubility of atorvastatin calcium, rosuvastatin calcium, simvastatin,pitavastatin calcium, and pravastatin sodium was evaluated in both aPUFA free acid solvent system (see Table 4) and a PUFA ethyl estersolvent system (Lovaza).

For this study, ten vials were prepared such that 5 vials contained 2 mLof Lovaza oil and 5 vials contained 2 mL of PUFA free acid solventsystem. About 20 mg of each statin were added to one Lovaza oil vial andto one PUFA free acid solvent system vial. Vials containing the sampleswere blanketed with nitrogen, then sealed. The vials were then sonicatedfor 1 hour or until contents appeared clear. After sonication, vialswhich remained clear received additional 20 mg aliquots of theirrespective statin and were again blanketed with nitrogen. The processwas repeated until all vials contained un-dissolved statin, with theexception of the pravastatin sodium in PUFA free acid solvent, which,after the addition of about 313 mg of pravastatin sodium, still remainedclear. The samples were allowed to sit overnight, after which allsamples were observed to contain un-dissolved statins. The samples werecentrifuged and the supernatant was assayed for the respective statin byHPLC. Assay results are shown in Table 7 below.

TABLE 7 Solubility in Lovaza (ethyl Solubility in PUFA ester) SolventFree Acid Solvent System System Drug Substance (mg/g) (mg/g)Atorvastatin Calcium 0.005 7.850 Rosuvastatin Calcium 0.001 12.700Simvastatin 19.267 45.783 Pitavastatin Calcium 0.615 12.771 PravastatinSodium 0.000 42.357

All of the tested statins displayed markedly higher solubility in thePUFA free fatty solvent system as compared to the PUFA ethyl estersolvent system.

Example 5 Stability Testing of Statins Dissolved in Omega-3 Free FattyAcids

The stability of atorvastatin calcium and rosuvastatin calciumsolubilized in a PUFA free acid solvent system was evaluated over thecourse of four weeks under 25° C., 60% relative humidity (RH) storageconditions.

Three batch formulations were prepared in parallel, by adding asufficient amount of PUFA free acid solvent system to (i) 2.0 g ofatorvastatin calcium, (ii) 2.0 g of rosuvastatin calcium, and (iii) nostatin (control), to fill a 200 mL vessel, followed by blanketing of thesolution with nitrogen and capping of the vessel. The compositions werethen mixed until full dissolution was observed, using a magnetic stirbar and sonication for 30-60 second intervals, taking care to maintain anitrogen headspace over the solution throughout processing.

Each statin-solvent system solution and the control solvent system wereeach respectively aliquoted into nine separate 20 mL Type I borosilicatevials, which were then blanketed with nitrogen, capped with aTeflon-coated serum stopper, and crimp sealed. Vials were covered withfoil to protect from light and stored at 25° C., 60% RH.

One vial of each of the three compositions was retrieved at eachtimepoint for testing. Procedures as listed in the USP Monograph forOmega-3-Acid Ethyl Ester Capsules were followed for measuring quantitiesof acid value, peroxide value, and absorbance for fats and fixed oils.The results are shown in Tables 8 and 9, below.

TABLE 8 Stability Testing for Atorvastatin Dissolved in PUFA free acidsolvent system t = 0 t = 1 week t = 2 week Statin in Oil mg/g Percentmg/g Percent mg/g Percent Atorvastatin Peak 1 3.659 41.87 1.632 19.721.509 19.25 Atorvastatin Peak 2 5.079 58.13 6.643 80.28 6.330 80.75Total 8.738 100 8.275 100 7.839 100

TABLE 9 Stability Testing for Atorvastatin Dissolved in PUFA free acidsolvent system t = 0 t = 1 week t = 2 week Statins in Oil mg/g Percentmg/g Percent mg/g Percent Rosuvastatin Peak 1 4.696 72.46 2.019 31.471.824 28.04 Rosuvastatin Peak 2 1.785 27.54 4.397 68.53 4.681 71.96Total 6.481 100 6.416 100 6.505 100

After pH adjustment, the two peaks were shown to reduce to a single peakof intact statin. No significant degradation was observed.

All publications, patents, patent applications and other documents citedin this application are hereby incorporated by reference in theirentireties for all purposes to the same extent as if each individualpublication, patent, patent application or other document wereindividually indicated to be incorporated by reference for all purposes.

While various specific embodiments have been illustrated and described,it will be appreciated that various changes can be made withoutdeparting from the spirit and scope of the invention(s).

What is claimed is:
 1. A pharmaceutical composition comprising a statindissolved in a PUFA free acid solvent system.
 2. The pharmaceuticalcomposition according to claim 1, wherein no more than about 10% byweight of the statin is undissolved in the solvent system.
 3. Thepharmaceutical composition according to claim 1, wherein the statin isdissolved in the PUFA free acid solvent system at a concentration of atleast about 5 mg statin per gram solvent system.
 4. The pharmaceuticalcomposition according to claim 3, wherein the statin is dissolved in thePUFA free acid solvent system at a concentration of at least about 7.5mg/g.
 5. The pharmaceutical composition according to claim 4, whereinthe statin is dissolved in the PUFA free acid solvent system at aconcentration of at least about 10 mg/g.
 6. The pharmaceuticalcomposition according to claim 5, wherein the statin is dissolved in thePUFA free acid solvent system at a concentration of at least about 20mg/g.
 7. The pharmaceutical composition according to claim 1, whereinthe statin is selected from the group consisting of pravastatin,lovastatin, simvastatin, atorvastatin, fluvastatin, rosuvastatin, andpitavastatin.
 8. The pharmaceutical composition according to claim 7,wherein the statin is rosuvastatin.
 9. The pharmaceutical compositionaccording to claim 7, wherein the statin is atorvastatin.
 10. Thepharmaceutical composition according to claim 7, wherein the statin issimvastatin.
 11. The pharmaceutical composition according to claim 1,wherein the PUFA free acid solvent system comprises EPA.
 12. Thepharmaceutical composition according to claim 11, wherein EPA is presentin the PUFA free acid solvent system in an amount of at least about 40%(a/a).
 13. The pharmaceutical composition according to claim 12, whereinDHA is present in the PUFA free acid solvent system in an amount of atleast about 13% (a/a).
 14. The pharmaceutical composition according toclaim 13, wherein DPA (C22:5 n-3) is present in the PUFA free acidsolvent system in an amount of at least about 1% (a/a).
 15. Thepharmaceutical composition according to claim 14, wherein the PUFA freeacid solvent system comprises about 45% (a/a) to about 65% (a/a) EPA,about 15% (a/a) to about 25% (a/a) DHA, and about 1% (a/a) to about 8%(a/a) DPA (C22:5 n-3).
 16. The pharmaceutical composition according toclaim 1, wherein the PUFA free acid solvent system comprises no morethan about 10% (a/a) omega-6 fatty acids in free acid form.
 17. An oralunit dosage form, wherein the dosage form comprises a capsule, thecapsule containing a pharmaceutical composition according to claim 1.18. The oral unit dosage form according to claim 17, wherein the capsulecomprises gelatin.
 19. The oral unit dosage form according to claim 18,wherein the capsule is a soft gelatin capsule.
 20. The oral unit dosageform of claim 19, wherein the soft gelatin capsule comprises Type Agelatin.
 21. The oral unit dosage form of claim 20, wherein at leastabout 1% (w/w) of the gelatin is Type A gelatin.
 22. The oral unitdosage form of claim 21, wherein the type A gelatin is porcine type Agelatin.
 23. The oral unit dosage form of claim 17, further comprising acoating on the exterior of the capsule.
 24. The oral unit dosage formaccording to claim 23, wherein the coating is a pH-sensitive entericcoating.
 25. The oral unit dosage form according to claim 23, whereinthe coating is a pH-insensitive coating.
 26. The oral unit dosage formaccording to claim 25, wherein the coating is apoly(ethylacrylate-methylacrylate) copolymer.
 27. The oral unit dosageform according to claim 26, wherein the coating is Eudragit NE-30 D. 28.The oral unit dosage form according to claim 17, wherein the capsulecontains about 1 to about 100 mg of a statin dissolved in about 500 mgto about 1 g of a PUFA free acid solvent system.
 29. The oral unitdosage form according to claim 28, wherein the capsule contains about2.5 mg to about 40 mg of a statin dissolved in about 1 g of a PUFA freeacid solvent system.
 30. The oral unit dosage form of claim 17, whereinthe statin is rosuvastatin.
 31. The oral unit dosage form of claim 17,wherein the statin is atorvastatin.
 32. The oral unit dosage form ofclaim 17, wherein the statin is simvastatin.
 33. A method of treating ablood lipid disorder, comprising administering a pharmaceuticalcomposition according to claim 1 to a subject with a blood lipiddisorder in an amount and for a duration sufficient to treat the bloodlipid disorder.
 34. The method according to claim 33, wherein the bloodlipid disorder is selected from the group consisting of:hypertriglyceridemia, hyperlipidemia, hypercholesterolemia, mixeddyslipidemia, and atherosclerosis.
 35. The method according to claim 33,wherein the method comprises administering at least 2 g of thepharmaceutical composition daily.
 36. The method according to claim 35,wherein the method comprises administering at least about 10 mg ofstatin daily.
 37. The method according to claim 33, wherein the PUFAfree acid solvent system of the pharmaceutical composition comprises atleast about 45% (a/a) EPA.
 38. The method according to claim 37, whereinthe statin in the pharmaceutical composition is rosuvastatin.
 39. Themethod according to claim 37, wherein the statin in the pharmaceuticalcomposition is atorvastatin.
 40. The method according to claim 37,wherein the statin in the pharmaceutical composition is simvastatin.